Our competitive renewal application addresses dry eye from the perspective of the new ocular-specific tear factor 'lacritin'discovered by us. Lacritin flows onto the ocular surface from lacrimal and meibomian glands to help promote ocular surface wetting. It is also a presumed contributor to corneal renewal via its basal corneal epithelial expression. Selective downregulation of lacritin in tears is associated with blepharitis and contact lens-related dry eye, the only two dry eye syndromes proteomically examined to date. Adding lacritin to eyes of normal rabbits or dry eye female monkeys ovariectomized one to two years earlier triggers elevated and sustained tear flow. These observations are reinforced by human corneal epithelial cell culture in which lacritin stimulates MUC16 protein production more efficiently than serum and by our original observation that lacritin augments tear peroxidase release from cultured rat lacrimal acinar cells independent of the parasympathetic- (carbachol-) stimulation pathway. Also subconfluent cultures of human corneal epithelial cells proliferate in a lacritin domain- and dose-specific manner. How lacritin promotes tearing and renewal is not understood. Our lacritin structure/function and lacritin signaling studies have sketched out a heparanase-dependent mechanism for targeting cell surface proteoglycan syndecan-1. This leads to putative ligation of a G-protein coupled receptor for signaling through G1i or G1o to PLC-PKC1/PLD/mTOR and PLC- PKC1/STIM1/calcineurin/Ca2+/ NFATC1. Differential upregulation of newly identified splice variants lacritin-b or -c (suspected to be incapable of respectively binding the GPCR or syndecan-1) coincident with downregulation of native lacritin, or loss of heparanase (observed preliminarily in dry eye tears) are two potential dry eye-provoking scenarios. Our working hypothesis is that lacritin is a regulator of ocular surface wetting and renewal. Our immediate goal is to characterize the signaling receptor and understand its mechanism of action. Our first aim is to understand specific protein-protein interactions constituting the lacritin/syndecan-1/G-protein coupled receptor heterocomplex. Our second aim is to characterize how upstream lacritin signaling is generated. Our third aim asks how downstream lacritin signaling can promote wetting and renewal towards a global appreciation for lacritin's role on the ocular surface. PUBLIC HEALTH RELEVANCE. Dry eye is very common. This project will work out how a new potential therapeutic for dry eye promotes ocular surface wetting. The therapeutic is based on a natural human tear protein.